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Suggestive evidence for association of two potassium channel genes with different idiopathic generalised epilepsy syndromes.

Several potassium channel genes have been implicated in epilepsy. We have investigated three such genes, KCNJ3, KCNJ6 and KCNQ2, by association studies using a broad sample of idiopathic generalised epilepsy (IGE) unselected by syndrome. One of the two single nucleotide polymorphisms (SNPs) examined in one of the inward rectifying potassium channel genes, KCNJ3, was associated with IGE by genotype (P=0.0097), while its association by allele was of borderline significance (P=0.051). Analysis of the different clinical subgroups within the IGE sample showed more significant association with the presence of absence seizures (P=0.0041) and which is still significant after correction for multiple testing. Neither SNP in the other rectifying potassium channel gene, KCNJ6, was associated with IGE or any subgroup. None of the three SNPs in the voltage-gated potassium channel gene, KCNQ2, was associated with IGE. However, one SNP was associated with epilepsy with generalised tonic clonic seizures only (P=0.016), as was an SNP approximately 56 kb distant in the closely linked nicotinic acetylcholine gene CHRNA4 (P=0.014). These two SNPs were not in linkage disequilibrium with each other, suggesting that if they are not true associations they have independently occurred by chance. Neither association remains significant after correcting for multiple testing.

3' Untranslated Regions↗

Predictive testing for multiple endocrine neoplasia type 1 using DNA polymorphisms.

Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominantly inherited predisposition to neoplastic lesions of the parathyroids, pancreas, and the pituitary. We have previously located the predisposing genetic defect to the long arm of chromosome 11 by genetic linkage. In this study, 124 members of six MEN1 families, including 59 affected individuals, were genotyped for restriction fragment length polymorphisms with different DNA probes, and the genetic linkage between these marker systems and MEN1 was determined. 13 marker systems (17 DNA probes) were found to be linked to MEN1. These markers are located within a region on chromosome 11 spanning 14% meiotic recombinations, with the MEN1 locus in the middle. Four of the marker systems are on the centromeric side of MEN1, and four on the telomeric side, based on meiotic crossovers. The remaining five DNA probes are closely linked to MEN1, with no crossovers in our set of families. The 13 marker systems can be used for an accurate and reliable premorbid test for MEN1. In most clinical situations it is possible to identify a haplotype of this part of chromosome 11 with the mutant MEN1 allele in the middle. The calculated predictive accuracy is greater than 99.5% if three such marker systems are informative. Therefore, genetic linkage testing can be used for informed genetic counseling in MEN1 families, and to avoid unnecessary biochemical screening programs.

Chromosome Mapping↗

Use of a global test for multiple outcomes in stroke trials with application to the National Institute of Neurological Disorders and Stroke t-PA Stroke Trial.

BACKGROUND: The National Institute of Neurological Disorders and Stroke (NINDS) held a workshop on statistical approaches to analysis of acute stroke trials that have multiple pre-specified outcomes. An objective was to plan for statistical analysis of the NINDS t-PA Stroke Trial, a randomized, double-blind, placebo-controlled trial of recombinant tissue plasminogen activator (rt-PA) for patients with acute ischemic stroke. Treatment success was defined as a "consistent and persuasive difference" in the proportion of patients achieving favorable outcomes on the Barthel Index, Modified Rankin Scale, Glasgow Outcome Scale, and National Institutes of Health Stroke Scale. The Data and Safety Monitoring Committee for the trial recommended this outcome because the committee did not believe that a positive result for a single outcome would provide sufficient evidence of efficacy. SUMMARY OF COMMENT: Workshop participants accepted the global test as a viable approach to testing the primary trial hypothesis. Clinician participants advocated categorizing outcomes as favorable/unfavorable, outcomes more clinically meaningful than continuous outcomes for evaluating a drug with potentially serious side effects. They agreed that a global test was appropriate for ischemic stroke when no single outcome is accepted. Hypothetical, special-case examples illustrate that highly correlated outcomes diminish the power of the global test. NINDS t-PA Stroke Trial data demonstrate the clinical interpretability of the global test. CONCLUSIONS: Workshop participants concluded that a global statistic should be used to test the trial's primary hypothesis accompanied by secondary tests of individual outcomes. Workshop participants recommended familiarizing the clinical/scientific community with the global approach.

Acute Disease↗

Association of polymorphism in glutathione S-transferase loci with susceptibility and outcome in rheumatoid arthritis: comparison with the shared epitope.

OBJECTIVE: To determine whether glutathione S-transferase GSTM1, GSTM3, GSTT1, and GSTP1 genotypes influence susceptibility or outcome in rheumatoid arthritis (RA). METHODS: 277 RA patients were compared with 577 controls to examine any associations between GST genotypes and susceptibility to RA. The effect of genotypes on outcome (Larsen and functional scores) and time integrated acute phase responses (erythrocyte sedimentation rate and C reactive protein) was assessed in 122 patients with disease duration of 5-10 years. GST and HLA-DRB1 genotypes were determined using polymerase chain reaction based assays. Data were analysed using multiple regression analysis with correction for age, sex, disease duration, and the DRB1 associated shared epitope (SE) and rheumatoid factor (RF) positivity where appropriate. RESULTS: The GSTM1*A/*B genotype was less common in RA cases (3 of 276) than in controls (22 of 591) (exact p = 0.047), though significance was lost when adjustment was made for multiple comparisons. The Larsen score was higher (p = 0.039) in the GSTM1 null patients (89.9) than those with other GSTM1 genotypes (74.7), and this was independent of the SE. Again, correction for multiple testing resulted in loss of significance. The difference in Larsen scores between patients homozygous or negative for the SE (87.9 v 74.3) was similar to that between GSTM1 null and non-null patients. No associations between GSTM3 or GSTT1 genotypes and disease markers were identified although the association between GSTP1*B/*B and Larsen score approached significance (p = 0.096). CONCLUSION: It is proposed that certain GSTs may influence susceptibility and radiological progression in RA and that this is independent of the effect of the HLA-DRB1 associated SE. The mechanism for this effect is presumed to be because of differences in the ability of various GST enzymes to utilise the cytotoxic products of oxidant stress. Although significance was lost after correction for multiple testing, the data indicate that further studies may be of value in RA to determine the influence of the GST and other genes involved in cellular protection against oxidative stress.

Acute-Phase Reaction↗

Reproducibility of low galactomannan enzyme immunoassay index values tested in multiple laboratories.

The Platelia galactomannan enzyme immunoassay is a commercially available nonculture method for diagnosing invasive aspergillosis. Recently, steps have been taken to improve sensitivity; specifically, a low (0.5 to 0.7) galactomannan index (GMI) value to determine positivity has been validated by multiple groups. We evaluated the intra-assay and interassay reproducibility at low index levels using three different kit lots on three different days in three different microbiology laboratories. Clinical and spiked sera were blinded and sent with galactomannan enzyme immunoassay (EIA) kits to the participating laboratories. We also prospectively collected data on all galactomannan EIAs performed between 1 September 2003 and 21 July 2004 at the University of Washington Medical Center microbiology laboratory to assess reproducibility of clinical samples analyzed in "real time." From the multilaboratory study, a total of 836 results were available for evaluation. Reproducibility was excellent between laboratories and on different days. Significant variability was seen between runs/lots, which may in part be associated with different threshold control values in different kits. Among the 1,410 clinical samples that were prospectively analyzed, 168 (90%) were confirmed to be positive on repeat testing (GMI, > or =0.5). Among the 19 (10.2%) initially positive samples not confirmed on repeat testing, the majority had a GMI at the threshold of the assay (between 0.5 and 0.7). Our findings suggest that the Platelia galactomannan immunoassay has good reproducibility. However, changes in GMI levels when different kit lots are used, and single samples with low-positive (GMI of 0.5 to 0.7) indices, should be interpreted with caution.

Antigens, Fungal↗

Diagnostic efficiency of neuropsychological test scores for discriminating boys with and without attention deficit-hyperactivity disorder.

A growing literature has documented group differences between boys with and without attention deficit-hyperactivity disorder (ADHD) on neuropsychological tests; however, whether or not such tests can discriminate individuals with ADHD from non-ADHD controls remains unclear. This study used conditional probability and receiver operating characteristic analyses to examine the efficiency of test-based diagnostic discriminations in a large sample of referred boys with and without ADHD. Single neuropsychological tests had limited discriminating ability at various cutoff scores. When multiple tests were used together, prediction of ADHD status improved but overall diagnostic efficiency remained limited. Diagnostic efficiency did not differ when medicated and nonmedicated index children were considered separately. Results suggest that children with ADHD show variable deficits on neuropsychological tests of attention and executive functions. Impairments on multiple neuropsychological tests are predictive of ADHD, but normal scores do not rule out the diagnosis. The prognostic implications of variable neuropsychological deficits in children with ADHD require further investigation.

Adolescent↗

Investigating users' requirements: computer-based anatomy learning modules for multiple user test beds.

OBJECTIVE: User data and information about anatomy education were used to guide development of a learning environment that is efficient and effective. The research question focused on how to design instructional software suitable for the educational goals of different groups of users of the Visible Human data set. The ultimate goal of the study was to provide options for students and teachers to use different anatomy learning modules corresponding to key topics, for course work and professional training. DESIGN: The research used both qualitative and quantitative methods. It was driven by the belief that good instructional design must address learning context information and pedagogic content information. The data collection emphasized measurement of users' perspectives, experience, and demands in anatomy learning. MEASUREMENT: Users' requirements elicited from 12 focus groups were combined and rated by 11 researchers. Collective data were sorted and analyzed by use of multidimensional scaling and cluster analysis. RESULTS: A set of functions and features in high demand across all groups of users was suggested by the results. However, several subgroups of users shared distinct demands. The design of the learning modules will encompass both unified core components and user-specific applications. The design templates will allow sufficient flexibility for dynamic insertion of different learning applications for different users. CONCLUSION: This study describes how users' requirements, associated with users' learning experiences, were systematically collected and analyzed and then transformed into guidelines informing the iterative design of multiple learning modules. Information about learning challenges and processes was gathered to define essential anatomy teaching strategies. A prototype instrument to design and polish the Visible Human user interface system is currently being developed using ideas and feedback from users.

Anatomy↗

[Epoxy resin as a cause of contact allergic eczema].

Epoxy resin-containing products of the plastic industry are increasingly employed. We retrospectively studied the incidence and course of epoxy resin contact dermatitis in 2265 patients in whom contact dermatitis was confirmed by patch testing. A positive patch test for epoxy resin was present in 31 patients (1.4%). Exposure to epoxy resin as the cause of contact dermatitis was 'certain' in 10, 'very likely' in 9, 'possible' in 4 and 'unclear' in 8 patients. Exposure to epoxy resin was related to occupational (55%) and leisure time activities (13%). In 48% patch tests were positive for multiple test allergens. Hands (56%), arms (38%) and face (38%) were mainly affected. Re-examination of 26 of 31 patients showed that a favourable course depended on identification of the cause of the exposure to epoxy resin, avoidance of further contact and absence of sensitization to multiple allergens.

Cross Reactions↗

Effect of diagnostic testing for multiple sclerosis on patient health perceptions. Rochester-Toronto MRI Study Group.

BACKGROUND AND METHODS: We examined the effect on health perceptions of a diagnostic workup in 172 patients with suspected multiple sclerosis (MS) using two questionnaires (Mishel, Rand Medical Outcome Study) and three visual analogue scales measuring distress. Patients were categorized into "MS," "not MS," or "still uncertain" groups. RESULTS: Patients with a definitive diagnosis reported a significant decrease in uncertainty (analysis of variance [ANOVA] P = .0004). The Medical Outcome Study showed a slightly worsened perceived current health status (ANOVA P = .02) and future health outlook (ANOVA P = .001) in the MS group compared with the other two groups. Distress over physical symptoms decreased in all three groups and anxiety was more likely to decrease than increase even in those with MS. Prognostic uncertainty distress decreased in the not MS group and still uncertain groups, whereas it remained unchanged in the MS group (P = .9156). CONCLUSION: Significant and generally beneficial changes in patient health perceptions are seen associated with a neurologic workup in suspected MS, irrespective of the final diagnosis.

Adolescent↗

Method and computer program for controlling the family-wise alpha rate in gene association studies involving multiple phenotypes.

Multiple significance testing involving multiple phenotypes is not uncommon in the context of gene association studies but has remained largely unaddressed. If no adjustment is made for the multiple tests conducted, the type I error probability will exceed the nominal (per test) alpha level. Nevertheless, many investigators do not implement such adjustments. This may, in part, be because most available methods for adjusting the alpha rate either: 1) do not take the correlation structure among the variables into account and, therefore, tend to be overly stringent; or 2) do not allow statements to be made about specific variables but only about multivariate composites of variables. In this paper we develop a simulation-based method and computer program that holds the actual alpha rate to the nominal alpha rate but takes the correlation structure into account. We show that this method is more powerful than several common alternative approaches and that this power advantage increases as the number of variables and their intercorrelations increase. The method appears robust to marked non-normality and variance heterogeneity even with unequal numbers of subjects in each group. The fact that gene association studies with biallelic loci will have (at most) three groups (i.e., AA, Aa, aa) implies by the closure principle that, after detection of a significant result for a specific variable, pairwise comparisons for that variable can be conducted without further adjustment of the alpha level.

Analysis of Variance↗

Genome-wide transcriptional response of a Saccharomyces cerevisiae strain with an altered redox metabolism.

The genome-wide transcriptional response of a Saccharomyces cerevisiae strain deleted in GDH1 that encodes a NADP(+)-dependent glutamate dehydrogenase was compared to a wild-type strain under anaerobic steady-state conditions. The GDH1-deleted strain has a significantly reduced NADPH requirement, and therefore, an altered redox metabolism. Identification of genes with significantly changed expression using a t-test and a Bonferroni correction yielded only 16 transcripts when accepting two false-positives, and 7 of these were Open Reading Frames (ORFs) with unknown function. Among the 16 transcripts the only one with a direct link to redox metabolism was GND1, encoding phosphogluconate dehydrogenase. To extract additional information we analyzed the transcription data for a gene subset consisting of all known genes encoding metabolic enzymes that use NAD(+) or NADP(+). The subset was analyzed for genes with significantly changed expression again with a t-test and correction for multiple testing. This approach was found to enrich the analysis since GND1, ZWF1 and ALD6, encoding the most important enzymes for regeneration of NADPH under anaerobic conditions, were down-regulated together with eight other genes encoding NADP(H)-dependent enzymes. This indicates a possible common redox-dependent regulation of these genes. Furthermore, we showed that it might be necessary to analyze the expression of a subset of genes to extract all available information from global transcription analysis.

Gene Deletion↗

Exercise time: a possible source of misleading results during long-term pharmacological studies by multiple stress testings in coronary artery disease.

The assessment of chronic pharmacological treatment of stable angina requires serial exercise stress testings. It is well known that exercise tolerance can be improved by the training effect of performing repeated testings. Our study investigated the values of heart rate, systolic blood pressure, rate-pressure product, and duration of exercise at 0.1 mV ST depression during exercise and the same parameters plus the maximal ST-segment depression at peak exercise, collected from three different tests. The first and second were performed at one-week intervals before, and the third (75 days after the first), was performed after a double-blind study with a drug versus placebo. We found a significant increase of exercise duration at 0.1 mV ST depression and at peak exercise, while 6 of 12 patients increased exercise duration from the second to the third test. Individual variability of exercise duration showed increasing values, ranging from 0 to 71% (first vs. third test). In contrast, the ratio of heart rate and systolic blood pressure did not differ between the tests. Our data criticized the use of mean values of exercise time for pharmacological studies; moreover, individual variability could affect results independently of drug or placebo administration. These findings should be taken into account in order to exclude misleading results.

Adrenergic beta-Antagonists↗

Genotoxic effects of p-aminophenol in Chinese hamster ovary and mouse lymphoma cells: results of a multiple endpoint test.

p-Aminophenol (PAP), a metabolite of aniline and acetaminophen, has been reported to be mutagenic in the L5178Y mouse lymphoma assay, but not in the CHO/HGPRT assay. In the present study, the effects of PAP in these two cells lines were examined to determine if the difference in activity is related to an intrinsic difference in the cell lines. CHO and L5178Y tk +/- mouse lymphoma cells were treated with PAP for 4 hr and assayed for 3 genetic endpoints: gene mutation at the HGPRT or TK locus, respectively; chromosomal aberrations at approximately 20 hr after initiation of treatment; and single-strand DNA breaks as detected by the single cell electrophoresis assay immediately after treatment. All treatments were conducted in the absence of S9. There was a dose-related, significant increase in TFT-resistant mouse lymphoma cells at dose levels that reduced survival to < or = 50% of concurrent controls. In CHO cells, however, there was no increase in thioguanine-resistant cells at dose levels that reduced cell survival to < 20%. These results are consistent with published reports on PAP. While the CHO cells were slightly more resistant to the toxic effects of PAP, the dose levels used in the two cell lines did not differ by more than 2-fold. At equivalent survival levels, PAP induced a significant (up to 20% aberrant cells) number of aberrations, primarily complex rearrangements, in both cell lines. In the single cell electrophoresis assay, there was a reproducible dose-related increase in cells with single-strand DNA breaks with both the L5178Y cells and the CHO cells. The induction of single-strand breaks and chromosome aberrations by PAP suggests that, mechanistically, PAP produces similar genetic damage in both CHO and L5178Y cell lines, but intrinsic differences between assay systems are responsible for the divergent gene mutation results.

Aminophenols↗